The present invention relates generally to the field of nucleic acid hybridization assays for detecting specific polynucleotide sequences and, more particularly, to compositions and methods for covalently attaching activated oligonucleotides to polymer-coated beads serving as solid supports.
Nucleic acid hybridization is a known method for identifying specific sequences of nucleic acids. Hybridization is based upon base pairing between complementary nucleic acid strands. When single stranded nucleic acids are incubated in appropriate buffer solutions, complementary base sequences pair to form double stranded stable molecules. The presence or absence of such pairing may be detected by several different methods described in the art.
Many known hybridization assays involve multiple steps, for example the hybridization technique described by Dunn, et al., Cell 12:23-36 (1977) (incorporated herein by reference), wherein a sandwich-type assay consists of a first hybridization between a "target" nucleic acid and a "capture" nucleic acid probe that has been immobilized on a solid support and a second hybridization between a "signal" nucleic acid probe, typically labeled with a radioactive isotope, and a different region of the immobilized target nucleic acid. The hybridization of the signal probe may then be detected by, for example, autoradiography.
Ranki, et al., U.S. Pat. No. 4,486,539 and U.S. Pat. No. 4,563,419 (both patents incorporated herein by reference), describe sandwich-type assays that first require steps to render nucleic acids single stranded before the single stranded nucleic acids are allowed to hybridize with a nucleic acid affixed to a solid carrier and with a nucleic acid labeled with a radioisotope.
Carrico, et al., U.S. Pat. No. 4,806,546, and Carrico, et al., European Patent Application 86112899.9 (both incorporated herein by reference), have described treatment of a nylon support with an alkylating agent to introduce amidine groups onto the surface of the nylon. The derived nylon surface possesses the capacity to noncovalently bind single stranded nucleic acids. The noncovalently bound nucleic acids are then used as probes to detect specific target nucleic acids in solution.
Hunger, et. al., Analytical Biochemistry 165:45-55 (1987); Hunger, et al., Analytical Biochemistry 156:286-299 (1986); Hunger, et al., European Patent Application 84109485.7 (all incorporated herein by reference), describe the use of cyanuric chloride-activated cellulose paper having immobilized restricted genomic DNA in Southern blot techniques for the detection of subpicogram quantities of complementary DNA. Biagioni, et. al., Analytical Biochemistry 89:616-619 (1978) (incorporated herein by reference), describe a method for the preparation of DNA-cellulose using cyanuric chloride wherein the DNA-cellulose is employed in affinity chromatography procedures.
Herzberg, et al., European Patent Application 0171150 (incorporated herein by reference), describe the use of oligonucleotides immobilized onto solid supports in dipstick assays.
Litman, et al., U.S. Pat. No. 4,391,904 (incorporated herein by reference, describes test strip kits wherein a member of an immunological pair is bonded to a solid surface. Also, Miller, et. al, Clin. Chem. 30:1467-1472 (1984), and Brown, et. al, Clin. Chem. 31:1500-1505 (1985) (both are incorporated herein by reference), describe an analytical test chamber containing cellulose threads coupled to an antibody as a solid matrix that permits multiple test results from a single sample.